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Cloning of Human Interleukin-2 cDNA in E. coli by Using Oligonucleotide Primers
강성만,김성완,정일엽,나도선,김지영,한문희,Kang, Seong-Man,Kim, Sung-Wan,Chung, Il-Yub,Na, Doe-Sun,Kim, Ji-Young,Han, Moon-Hi 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.1
사람 인터루킨-2(IL-2)의 cDNA 클론을 oligo-누클레오티드를 primer로 사용하여 분리하였다. 사람 leukaemic T-cell line인 Jurkat 세포로부터 mRNA를 분리 하였다. ss-cDNA를 합성하기 위하여 인터루킨-2를 코딩하는 mRNA의 3' 끝쪽에 상보적인 30 mer oligo-누클레오티드를 역전사 반응시 primer로 사용하였으며, ds-cDNA를 합성하기 위해서는 만들어진 ss-cDNA의 3' 끝쪽에 상보적인 oligo-누클레오티드를 primer로 사용하였다. 이 ds-cDNA를 사용하여 partial cDNA library를 만든 뒤 cDNA 합성에 사용한 oligo-누클레오티드를 probe로 사용하여 콜로니 hybridization을 하여 인터루킨-2 cDNA를 찾기위하여 screen하였다. 약 200개의 transformants 중에서 세 클론이 positive signal을 나타냈다. 제한효소지도를 작성하고 누클레오티드 염기서열을 결정함으로써 이들 세 클론이 모두 인터루킨-2 cDNA를 포함하고 있음을 밝혔다. 이 결과는 우리가 만든 partial cDNA library에 인터루킨-2 cDNA가 Taniguchi 등 (1983)이 만든 total cDNA library에 들어있는 것보다 약 300배 가량 증가되어 있음을 시사한다. A cDNA clone for human interleukin-2 (IL-2) was isolated by using oligonucleotides as primers for the first and the second cDNA syntheses. Total RNA was prepared from Jurkat, a human leukaemic T-cell line, cells and mRNA was isolated. To synthesize ss-cDNA, a 30 mer oligonucleotide was used as a primer in the reverse transcriptase reaction. The sequence of the oligonucleotide was complementary to the 3' end of the coding sequence of IL-2. ds-cDNA was synthesized by DNA polymerase reaction using another oligonucleotide as a primer. A partial cDNA library was prepared using the ds-cDNA and screened for the presence of IL-2 cDNA by colony hybridization using the same oligonucleotides that were used in the cDNA synthesis reactions as probes. Three out of 200 transformants showed positive signals. Analysis of these three clones by restriction enzyme mapping and nucleotide sequencing showed that all of them contained IL-2 cDNA. Our results indicated that the IL-2 cDNA was enriched in the partial cDNA library about 300 fold over the population of IL-2 cDNA in the total cDNA library reported by Taniguchi et al. (1983).
Oligo - 누클레오티드 primer 를 이용한 사람 인터루킨 - 2 - cDNA의 E . coli 내 클로닝
강성만,김성완,정일엽,나도선,김지영,한문희 ( Seong Man Kang,Sung Wan Kim,Il Yub Chung,Doe Sun Na ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.1
A cDNA clone for human interleukin-2 (IL-2) was isolated by using oligonucleotides as primers for the first and the second cDNA syntheses. Total RNA was prepared from Jurkat, a human leukaemic T-cell line, cells and mRNA was isolated. To synthesize ss-cDNA, a 30 mer oligonucleotide was used as a primer in the reverse transcriptase reaction. The sequence of the oligonucleotide was complementary to the 3` end of the coding sequence of IL-2. ds-cDNA was synthesized by DNA polymerase reaction using another oligonucleotide as a primer. A partial cDNA library was prepared using the ds-cDNA and screened for the presence of IL-2 cDNA by colony hybridization using the same oligonucleotides that were used in the cDNA synthesis reactions as probes. Three out of 200 transformants showed positive signals. Analysis of these three clones by restriction enzyme mapping and nucleotide sequencing showed that all of them contained IL-2 cDNA. Our results indicated that the IL-2 cDNA was enriched in the partial cDNA library about 300 fold over the population of IL-2 cDNA in the total cDNA library reported by Taniguchi et al. (1983).
Seong Jun Lim,Hee Jeong Kim,Il Young Chung,Ji Sun Kim,Sae Byul Lee,Jong Won Lee,Byung Ho Son,Sei Hyun Ahn,Hak Hee Kim,Sung-Bae Kim,Gyung-Yub Gong,고범석 한국유방암학회 2020 Journal of Breast Disease Vol.8 No.1
Purpose: Various incision techniques have been used in breast-conserving surgery (BCS) to reduce scar formation, including the frequently used periareolar incision technique. However, its applicability in removing large-size tumors or those distant from the nipple has remained limited. We performed periareolar zigzag (P-Z) incision to address these problems in BCS and to improve cosmetic outcomes. Methods: Patients who underwent P-Z incision in BCS between January 2016 and November 2017 were retrospectively analyzed in terms of clinicopathological features and surgical findings. Factors affecting the positive margin were analyzed. Results: A total of 305 patients were reviewed. The patients presented with a median age of 51 years, mean tumor size 1.9 cm, and mean tumor distance of 3.3 cm from the nipple. Intraoperative frozen biopsy and final pathologic findings showed 43 (14.1%) and 7 (2.3%) tumor-positive margins. There were no major complications associated with the surgery, and no re-operations were required due to bleeding or other reasons. Conclusion: The P-Z incision technique was used to achieve sufficient operative field during BCS, and large tumors or those distant from the nipple could be removed without any complications. This technique is considered an oncologically safe surgical technique resulting in good cosmetic outcomes.
Oh, Joo Han,Yoon, Jong Pil,Kim, Dong Hyun,Chung, Seok Won,Kim, Joon Yub,Lee, Hyun-Joo,Il, Seo,Park, Kyeong Hyeon,Lee, Hoseok Elsevier 2019 Journal of shoulder and elbow surgery Vol.28 No.10
<P><B>Background</B></P> <P>The correlation between shoulder strength deficits and function in rotator cuff tears remains uncertain. This study aimed to determine the correlation between shoulder strength deficits and shoulder function evaluated by various clinical scoring systems.</P> <P><B>Methods</B></P> <P>A total of 262 patients (mean age, 59.67 years [standard deviation, 8.06 years]) who underwent full-thickness rotator cuff repair were included. Patients in group I (n = 188) had small to large rotator cuff tears, whereas those in group II (n = 74) had massive rotator cuff tears. Demographic factors, isokinetic test results, and shoulder function evaluated using various scoring systems were obtained. Correlation differences according to severity of the rotator cuff tear were evaluated.</P> <P><B>Results</B></P> <P>We found weak correlations between shoulder strength deficits (peak torque and total work) and clinical outcomes in patients with rotator cuff tears (<I>r</I> = –0.288). For patients in group I (nonmassive tears), we found a weaker correlation (<I>r</I> = –0.242) according to the tear pattern. However, shoulder strength deficits in group II patients (massive tears) were strongly correlated with American Shoulder and Elbow Surgeons (<I>r</I> = –0.598), Constant (<I>r</I> = –0.582), and Short Form 36 (<I>r</I> = –0.511) scores, especially regarding internal rotator strength deficits.</P> <P><B>Conclusions</B></P> <P>Shoulder strength deficits measured via isokinetic testing and shoulder function were weakly correlated in patients with rotator cuff tears. However, shoulder strength deficits in patients with massive tears considerably worsened shoulder function and systemic disability, but not regional disability. In particular, internal rotator strength deficits were strongly correlated with poor shoulder function.</P>
( Hyun Ji Cho ),( Jeong Han Kang ),( Ji Hak Jeong ),( Yun Jeong Jeong ),( Kwan Kyu Park ),( Yoon Yub Park ),( Yong Suk Moon ),( Hong Tae Kim ),( Il Kyung Chung ),( Cheorl Ho Kim ),( Hyeun Wook Chang ) 영남대학교 약품개발연구소 2012 영남대학교 약품개발연구소 연구업적집 Vol.22 No.0
Fibrosis is induced by the excessive and abnormal deposition of extracellular matrix (ECM) with various growth factors in tissues. Transforming growth factor-β1 (TGF-β1), the growth factor involved in fibrosis, modulates ECM synthesis and accumulation. TGF-β1 enhances the production of stimulators of ECM synthesis such as plasminogen activator inhibitor type 1 (PAI-1). As such, PAI-1 expressiondirectly influences the proteolysis, invasion, and accumulation of ECM. It was shown in this study thatascochlorin, a prenylpenl antiobiotic, prevents the expression of profibrotic factors, such as PAI-1 and collagen type I, and that the TGF-β1-induced PAI-1 promoter activity is inhibited by ascochlorin.Ascochlorin abolishes the phosphorylation of the EGFR-MEK-ERK signaling pathway to regulate the TGF-β1-induced expression of PAI-1 without the inhibition of TβRII phosphorylation. Furthermore, the MEK inhibitor and EGFR siRNA block PAI-1 expression, and the Raf-1, MEK, and ERK signaling pathways for the regulation of PAI-1 expression. Ascochlorin suppresses the matrix metalloproteinases (MMPs) activity to activate the heparin-binding EGF-like growth factor (HB-EGF), to induce the phosphorylation of EGFR, and the MMPs inhibitor suppresses EGFR phosphorylation and the PAI-1 mRNA levels. These results suggest that ascochlorin prevents the expression of PAI-1via the inhibition of an EGFR-dependent signal transduction pathway activated by MMPs.
( Ji Hak Jeong ),( Yun Jeong Jeong ),( Hyun Ji Cho ),( Jae Moon Shin ),( Jeong Han Kang ),( Kwan Ku Park ),( Yoon Yub Park ),( Il Kyung Chung ),( Hyeun Wook Chang ),( Jun Ji Magae ),( Shin Sung Kang ) 영남대학교 약품개발연구소 2012 영남대학교 약품개발연구소 연구업적집 Vol.22 No.0
Ascochlorin, a non-toxic prenylphenol compound derived from the fungus Ascochyta viciae, has been shown recently to have anti-cancer effects on various human cancer cells. However, the precise molecular mechanism of this anti-cancer activity remains to be elucidated. Here, we investigated the effects of ascochlorin on hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) expression in human epidermoid cervical carcinoma CaSki cells. Ascochlorin inhibited epidermal growth factor (EGF)-induced HIF-1α and VEGF expression through multiple potential mechanisms. First, ascochlorin selectively inhibited HIF-1α expression in response to EGF stimulation, but not in response to hypoxia (1% O(2)) or treatment with a transition metal (CoCl(2)). Second, ascochlorin inhibited EGF-induced ERK-1/2 activation but not AKT activation, both of which play essential roles in EGF-induced HIF-1α protein synthesis. Targeted inhibition of epidermal growthfactor receptor (EGFR) expression using an EGFR-specific small interfering RNA (siRNA) diminished HIF-1α expression, which suggested that ascochlorin inhibits HIF-1α expression through suppression of EGFR activation. Finally, we showed that ascochlorin functionally abrogates in vivo tumor angiogenesis induced by EGF in a Matrigel plug assay. Our data suggest that ascochlorin inhibits EGF-mediated induction of HIF-1α expression in CaSki cells, providing a potentially new avenue of development of anti-cancer drugs that target tumor angiogenesis. 113: 1302-1313, 2012. ⓒ 2011 Wiley Periodicals. Inc.